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product-image-AAA127105_FACS8.jpg FCM/FACS (Flow Cytometry) (Figure 5. Flow Cytometry analysis of PC-3 cells using anti-CD276 antibody (AAA127105).Overlay histogram showing PC-3 cells stained with AAA127105 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CD276 Antibody (AAA127105, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)

Rabbit anti-Human CD276 Polyclonal Antibody | anti-CD276 antibody

Anti-CD276 Antibody Picoband

Gene Names
CD276; B7H3; B7-H3; B7RP-2; 4Ig-B7-H3
Reactivity
Human
Applications
ELISA, Flow Cytometry, Functional Assay, Immunohistochemistry, Western Blot
Purity
Immunogen affinity purified.
Synonyms
CD276, Antibody; Anti-CD276 Antibody Picoband; Cone-rod homeobox protein; colony stimulating factor 2 receptor, beta, low-affinity (granulocyte-macrophage); Cytokine receptor common subunit beta; GM-CSF/IL-3/IL-5 receptor common beta subunit; CD131; Csf2rb; Aic2b; Csf2rb1; Il3rb1; anti-CD276 antibody
Ordering
Host
Rabbit
Reactivity
Human
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Broadly expressed with higher expression in heart.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Applicable Applications for anti-CD276 antibody
ELISA, FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot)
Immunogen
E coli-derived human CD276 recombinant protein (Position: L29-Q458).
Subcellular Localization
Membrane. Single-pass type I membrane protein
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500ug/ml.
Cross Reactivity
No cross-reactivity with other proteins.
Protein Function
High affinity receptor for interleukin-3, interleukin-5 and granulocyte-macrophage colony-stimulating factor.
Preparation and Storage
At -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months.Avoid repeated freezing and thawing.

FCM/FACS (Flow Cytometry)

(Figure 5. Flow Cytometry analysis of PC-3 cells using anti-CD276 antibody (AAA127105).Overlay histogram showing PC-3 cells stained with AAA127105 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CD276 Antibody (AAA127105, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)

product-image-AAA127105_FACS8.jpg FCM/FACS (Flow Cytometry) (Figure 5. Flow Cytometry analysis of PC-3 cells using anti-CD276 antibody (AAA127105).Overlay histogram showing PC-3 cells stained with AAA127105 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CD276 Antibody (AAA127105, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of CD276 using anti-CD276 antibody (AAA127105).CD276 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-CD276 Antibody (AAA127105) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

product-image-AAA127105_IHC10.jpg IHC (Immunohistochemistry) (Figure 4. IHC analysis of CD276 using anti-CD276 antibody (AAA127105).CD276 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-CD276 Antibody (AAA127105) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemisry)

(Figure 3. IHC analysis of CD276 using anti-CD276 antibody (AAA127105).CD276 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-CD276 Antibody (AAA127105) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

product-image-AAA127105_IHC11.jpg IHC (Immunohistochemisry) (Figure 3. IHC analysis of CD276 using anti-CD276 antibody (AAA127105).CD276 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-CD276 Antibody (AAA127105) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohiostchemistry)

(Figure 2. IHC analysis of CD276 using anti-CD276 antibody (AAA127105).CD276 was detected in a paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-CD276 Antibody (AAA127105) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

product-image-AAA127105_IHC13.jpg IHC (Immunohiostchemistry) (Figure 2. IHC analysis of CD276 using anti-CD276 antibody (AAA127105).CD276 was detected in a paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-CD276 Antibody (AAA127105) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of CD276 using anti-CD276 antibody (AAA127105).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human U2OS whole cell lysates,Lane 2: human PC-3 whole cell lysates,Lane 3: human Hela whole cell lysates,Lane 4: human MCF-7 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CD276 antigen affinity purified polyclonal antibody (#AAA127105) at 0.5ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CD276 at approximately 90-100 kDa. The expected band size for CD276 is at 100 kDa.)

product-image-AAA127105_WB15.jpg WB (Western Blot) (Figure 1. Western blot analysis of CD276 using anti-CD276 antibody (AAA127105).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human U2OS whole cell lysates,Lane 2: human PC-3 whole cell lysates,Lane 3: human Hela whole cell lysates,Lane 4: human MCF-7 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CD276 antigen affinity purified polyclonal antibody (#AAA127105) at 0.5ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CD276 at approximately 90-100 kDa. The expected band size for CD276 is at 100 kDa.)
Related Product Information for anti-CD276 antibody
The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
References
1. Chapoval, A. I., Ni, J., Lau, J. S., Wilcox, R. A., Flies, D. B., Liu, D., Dong, H., Sica, G. L., Zhu, G., Tamada, K., Chen, L. B7-H3: a costimulatory molecule for T cell activation and IFN-gamma production. Nature Immun. 2: 269-274, 2001. 2. Steinberger, P., Majdic, O., Derdak, S. V., Pfistershammer, K., Kirchberger, S., Klauser, C., Zlabinger, G., Pickl, W. F., Stockl, J., Knapp, W. Molecular characterization of human 4Ig-B7-H3, a member of the B7 family with four Ig-like domains. J. Immun. 172: 2352-2359, 2004. 3. Suh, W.-K., Gajewska, B. U., Okada, H., Gronski, M. A., Bertram, E. M., Dawicki, W., Duncan, G. S., Bukczynski, J., Plyte, S., Elia, A., Wakeham, A., Itie, A., and 12 others. The B7 family member B7-H3 preferentially down-regulates T helper type 1-mediated immune responses.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
57,235 Da
NCBI Official Full Name
B7-H3 protein
NCBI Official Synonym Full Names
CD276 molecule
NCBI Official Symbol
CD276
NCBI Official Synonym Symbols
B7H3; B7-H3; B7RP-2; 4Ig-B7-H3
NCBI Protein Information
CD276 antigen; B7 homolog 3; costimulatory molecule
UniProt Protein Name
CD276 antigen
UniProt Gene Name
CD276
UniProt Synonym Gene Names
B7H3; B7-H3
UniProt Entry Name
CD276_HUMAN

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Product Notes

The CD276 cd276 (Catalog #AAA127105) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-CD276 Antibody Picoband reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's CD276 can be used in a range of immunoassay formats including, but not limited to, ELISA, FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot). Researchers should empirically determine the suitability of the CD276 cd276 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CD276, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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